Hes-1在人脑胶质瘤干细胞分化过程中的动态表达及意义

The Expression and Significance of Hes-1 protein during human brain glioma stem cells differentiation

目的研究在体外培养的人脑胶质瘤干细胞诱导分化过程中Hes-1蛋白表达的动态变化。方法对体外培养的人脑胶质瘤干细胞进行诱导分化,于未分化、分化第3、7d收集标本,行Hes-1蛋白免疫荧光染色后流式细胞术检测标本中阳性细胞比例;Hes-1分别与CD133、GFAP、MAP2、MBP蛋白双标免疫荧光染色后,激光共聚焦显微镜下观察Hes-1蛋白表达强度及细胞类型的变化。结果随着人脑胶质瘤干细胞的分化,Hes-1蛋白阳性细胞比例由未分化至分化第3d、第7d有明显下降(P<0.01);Hes-1蛋白在CD133、MAP2、GFAP和MBP阳性细胞均可表达,但在较幼稚的细胞中表达较强,随着细胞分化成熟,其表达明显减弱,分化第7d时大部分分化成熟的肿瘤细胞(MAP2、GFAP和MBP阳性细胞)中表达极弱或不表达。结论随着人脑胶质瘤干细胞的分化,Hes-1蛋白表达的阳性细胞比例和强度均明显下降,提示Hes-1表达的动态变化可能与调控人脑胶质瘤干细胞的分化过程有关。

Objective To study the dynamic expression of Hes-1 protein in human brain glioma stem ceils （BGSCs） differentia- tion in vitro. Methods Cells were harvested at 0 day, the 3rd day and the 7th day after BGSCs differentiation, then the ratio of Hes- 1 positive cells were counted by flow cytometry, and the expression intensity and cell types were detected by laser coxffocal microscopy after immunofluorescence staining with Hes-1 and GFAP, MAP2 or MBP. Results The ratio of Hes-1 positive cells significantly de- creased at the 3rd day and the 7th day along with BGSCs differentiation （ P 〈0.01 ）. Hes-1 protein expressed in CD133, GFAP, MAP2 and MBP positive cells, and the expression level obviously decreased along with BGSCs differentiation. Hes-1 protein expressed intensively in inmlature cells, and weakly or negatively in mature cells. Conclusion The expression of Hes-1 protein significantly decreased along with BGSCs differentiation, which hints that have been detected in human gliomas and normal cerebral tissuses, and the expression level in ghomas, especially in those of grade Ⅲ - Ⅳ, was considerably higher than those in normal cerebral tissuses. Which hints that the dynamic expression of Hes-1 protein may take part in controlling the differentiation of BGSCs.